Heterocyclic compounds as kinase inhibitors for therapeutic uses

ABSTRACT

Heterocyclic compounds of formula I shown below and pharmaceutical compositions containing one of such compounds: Also disclosed is a method of treating a condition modulated by the colony-stimulating factor-1 receptor with one of the heterocyclic compounds.

BACKGROUND

Colony-stimulating factor-1 receptor (CSF1R) is a member of tyrosinekinase class III. It plays an important role in cell proliferation,differentiation, migration, and survival. See Cannarile et al., J.Immunother. Cancer, 2017, 5:53. Deregulation of this tyrosine kinase isassociated with various disorders and diseases, such as inflammatorydisorders, neurological disorders, cardiovascular diseases, bone-relateddiseases, and cancers.

Recent studies have shown that CSF1R is related to differentiation oftumor-associated macrophages (TAMs). See El-Gamal et al., J. Med. Chem.,2018, 61, 5450-5466. Specifically, TAMs express, on their surfaces,CSF1R, which forms a signaling axis with an active ligand, i.e., colonystimulating factor-1 (CSF1). When activated, the CSF1R/CSF1 signalingaxis promotes proliferation of monocytes, differentiation of themonocytes into TAMs, and survival of the TAMs.

Overexpression of CSF1 in several forms of cancer has been associatedwith activation and recruitment of TAMs to tumor sites. TAMs modifytumor microenvironment to render it more conducive to cancer cellgrowth, angiogenesis, and metastasis. Further, they can cause localizedimmunosuppression in tumor tissues, resulting in resistance to cancertherapy. As such, inhibiting the CSF1R/CSF1 signaling axis presents anattractive avenue for treating cancers associated with overexpression ofCSF1.

Thus, there is a need to provide compounds that selectively inhibitCSF1R, demonstrate favorable safety profiles, and also exhibit high invivo efficacy in treating cancers associated with CSF1R.

SUMMARY

The present invention is based on unexpected discoveries that certainheterocyclic compounds effectively inhibit colony-stimulating factor-1receptor (CSF1R).

In one aspect, this invention relates to these heterocyclic compoundsand other heterocyclic compounds analogous thereto covered by formula I:

in which A is H, C₁₋₆ alkyl, or ORr, Rr being H or C₁₋₆ alkyl; Y¹ isphenyl substituted with (R¹)_(n), 5-membered heteroaryl substituted with(R²)_(o), 5-membered heterocycloalkenyl substituted with (R²)_(o), oralkenyl substituted with (R²)_(o), in which R¹ in (R¹)_(n), n being 0-4,is, independently, F, Cl, Br, NO₂, CN, amino, C₁-C₆ alkyl, C₁-C₆haloalkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₈ cycloalkyl, C₅-C₁₅heterocycloalkyl, aryl, heteroaryl, carbonyl, thionyl, iminyl, orspiroamino; and R² in (R²)_(o), o being 0-5, is, independently, F, Cl,Br, NO₂, CN, amino, C₁-C₆ alkyl, C₁-C₆ haloalkyl, C₂-C₆ alkenyl, C₂-C₆alkynyl, C₃-C₈ cycloalkyl, C₅-C₁₅ heterocycloalkyl, aryl, heteroaryl,carbonyl, thionyl, iminyl, spiroamino, or C₁-C₆ alkoxyl; X¹ is N or CR³,R³ being H, F, Cl, Br, CN, C₁-C₆ alkyl, C₁-C₆ haloalkyl, or C₁-C₆alkoxyl; X² is O, S, NH, or CH₂; Y² is

in which each of Q¹, Q², Q³, Q⁴, Q⁵, Q⁶, Q⁷ and Q⁸ is, independently, Nor CR⁴, R⁴ being H, F, Cl, Br, CN, amino, C₁-C₆ alkyl, C₁-C₆ haloalkyl,or C₁-C₆ alkoxyl; Z¹ is O, S, or NRr; Z² is O, S, or NRr; and G and Hare, respectively, C or N and N or C; X³ is deleted, CH₂, (CH₂)₂, orCH(C≡CH); Y³ is C₁-C₆ alkyl, aryl, heteroaryl, C₃-C₈ cycloalkyl, orC₅-C₆ heterocycloalkyl having one heteroatom, in which the oneheteroatom is O or N; and X⁴ in (X⁴)_(m), m being 0-5, is,independently, F, Cl, Br, CN, SO₂NH₂, amino, C₁-C₆ alkyl, or C₁-C₆alkoxyl.

The term “alkyl” refers to a straight or branched monovalent hydrocarbonmoiety containing 1-20 carbon atoms, e.g., methyl, ethyl, n-propyl,i-propyl, n-butyl, i-butyl, and t-butyl. The term “haloalkyl” refers toan alkyl group substituted with one or more halogen atoms. The term“alkenyl” refers to a straight or branched monovalent or bivalenthydrocarbon containing 2-20 carbon atoms and one or more double bonds,e.g., ethenyl, propenyl, propenylene, allyl, and 1,4-butadienyl. Theterm “alkynyl” refers to a straight or branched monovalent or bivalenthydrocarbon containing 2-20 carbon atoms and one or more triple bonds,e.g., ethynyl, ethynylene, 1-propynyl, 1- and 2-butynyl, and1-methyl-2-butynyl. The term “aryl” refers to a monovalent 6-carbonmonocyclic, 10-carbon bicyclic, 14-carbon tricyclic aromatic ringsystem, e.g., phenyl, naphthyl, and anthracenyl. The term “heteroaryl”refers to a monovalent aromatic 5-8 membered monocyclic, 8-12 memberedbicyclic, or 11-14 membered tricyclic ring system having one or moreheteroatoms (such as O, N, S, or Se), e.g., imidazolyl, pyrazoyl,oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridyl, pyrazinyl,pyrimidyl, pyridazinyl, furyl, and thienyl. The term “cycloalkyl” refersto a monovalent or bivalent saturated hydrocarbon ring system having 3to 30 carbon atoms (e.g., C₃-C₁₂), e.g., cyclopropyl, cyclobutyl,cyclopentyl, cyclopentenyl, cyclohexyl, 1,4-cyclohexylene, cycloheptyl,and cyclooctyl. The term “cycloalkenyl” refers to a monovalent orbivalent non-aromatic hydrocarbon ring system having 3 to 30 carbons(e.g., C₃-C₁₂) and one or more double bonds, e.g., cyclopentenyl,cyclohexenyl, and cycloheptenyl. The term “heterocycloalkyl” refers to amonovalent or bivalent nonaromatic 5-8 membered monocyclic, 8-12membered bicyclic, or 11-14 membered tricyclic ring system having one ormore heteroatoms (such as O, N, S, or Se), e.g., piperidinyl,piperazinyl, pyrrolidinyl, dioxanyl, morpholinyl, tetrahydrofuranyl, andtetrahydropyranyl. The term “heterocycloalkenyl” refers to a monovalentor bivalent nonaromatic 5-8 membered monocyclic, 8-12 membered bicyclic,or 11-14 membered tricyclic ring system having one or more heteroatoms(such as O, N, S, or Se) and one or more double bonds. The term “amino”refers to a —NRR′ moiety, in which R and R′ are, independently, H,alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocycloalkyl,heterocycloalkenyl, aryl, heteroaryl, aralkyl, or heteroarakyl. The term“carbonyl” refers to a —C(O)R moiety, in which R is H, alkyl, alkenyl,alkynyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl,alkoxyl, amino, aryl, or heteroaryl. The term “thionyl” refers to a—S(O)R moiety, in which R is H, alkyl, alkenyl, alkynyl, cycloalkyl,cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, alkoxyl, amino,aryl, or heteroaryl. The term “iminyl” refers to a —C(NR)R′, in which Ris H or C₁-C₆ alkyl and R′ is H, alkyl, alkenyl, alkynyl, cycloalkyl,cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, alkoxyl, amino,aryl, or heteroaryl. The term “spiroamino” refers to a monovalent 7-11membered bicyclic spiro moiety containing one N or a monovalent 10-16membered tricyclic spiro moiety containing one N.

In addition, alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl,cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, and alkoxyl can besubstituted or unsubstituted. Possible substituents include, but are notlimited to, D, CN, NO₂, halo, C₁-C₂₀ alkyl, C₂-C₂₀ alkenyl, C₂-C₂₀alkynyl, C₁-C₁₀ alkoxyl, C₃-C₃₀ cycloalkyl, C₃-C₃₀ cycloalkenyl, C₃-C₃₀heterocycloalkyl, C₃-C₃₀ heterocycloalkenyl, aryl, aryloxyl, heteroaryl,heteroaryloxyl, amino, halo, oxo (O═), thioxo (S═), thio, silyl, C₁-C₁₀alkylthio, arylthio, C₁-C₁₀ alkylsulfonyl, arylsulfonyl, acylamino,aminoacyl, aminothioacyl, amidino, mercapto, amido, thioureido,thiocyanato, sulfonamido, guanidine, ureido, acyl, thioacyl, acyloxy,carbamido, carbamyl, carboxyl, and carboxylic ester.

The compounds of formula I include the compounds themselves, as well astheir salts, their stereoisomers, their solvates, their tautomers, theirdeuterated analogues, and their prodrugs, if applicable. A salt, forexample, can be formed between an anion and a positively charged group(e.g., ammonium) on a heterocyclic compound of this invention. Suitableanions include chloride, bromide, iodide, sulfate, bisulfate, sulfamate,nitrate, phosphate, citrate, methanesulfonate, trifluoroacetate,glutamate, glucuronate, glutarate, malate, maleate, succinate, fumarate,tartrate, tosylate, salicylate, lactate, naphthalenesulfonate, andacetate. Likewise, a salt can also be formed between a cation and anegatively charged group (e.g., carboxylate) on a heterocyclic compound.Suitable cations include sodium ion, potassium ion, magnesium ion,calcium ion, and an ammonium cation such as tetramethylammonium ion. Thesalts of the heterocyclic compounds of this invention can also containquaternary nitrogen atoms. Examples of prodrugs include esters and otherpharmaceutically acceptable derivatives, which, upon administration to asubject, are capable of providing active heterocyclic compounds. Asolvate refers to a complex formed between an active heterocycliccompound and a pharmaceutically acceptable solvent, e.g., water,ethanol, isopropanol, ethyl acetate, acetic acid, and ethanolamine.

An additional aspect of this invention relates to a pharmaceuticalcomposition containing one or more of the heterocyclic compounds coveredby formula I. The pharmaceutical composition can be used for treatmentof a CSF1R modulated condition.

Also within the scope of this invention is a method of treating a CSF1Rmodulated condition, e.g., a cancer, an inflammatory disorder, a bonedisorder, or an autoimmune disease. The method includes administering toa subject in need thereof an effective amount of one or more of theabove-described heterocyclic compounds.

The term “treatment” or “treating” refers to administering one or moreheterocyclic compounds of this invention to a subject who has a CSF1Rmodulated condition, a symptom of such a condition, or a predispositiontoward it, with the purpose of conferring a therapeutic or prophylacticeffect. “An effective amount” refers to the amount of an active compoundthat is required to confer such effect. Effective doses will vary, asrecognized by those skilled in the art, depending on the types ofdisease treated, route of administration, excipient usage, and thepossibility of co-usage with other therapeutic treatment.

A pharmaceutical composition of this invention can be administeredparenterally, orally, nasally, rectally, topically, or buccally. Theterm “parenteral” as used herein refers to subcutaneous, intracutaneous,intravenous, intraperitoneal, intramuscular, intraarticular,intraarterial, intrasynovial, intrasternal, intrathecal, intralesional,or intracranial injection, as well as any suitable infusion technique.

A sterile injectable composition can be a solution or suspension in anon-toxic parenterally acceptable diluent or solvent, such as a solutionin 1,3-butanediol. Among the acceptable vehicles and solvents that canbe employed are mannitol, water, Ringer's solution, and isotonic sodiumchloride solution. In addition, fixed oils are conventionally employedas a solvent or suspending medium (e.g., synthetic mono- ordi-glycerides). Fatty acids, such as oleic acid and its glyceridederivatives, are useful in preparation of injectables, as are naturalpharmaceutically acceptable oils, such as olive oil and castor oil,especially in their polyoxyethylated versions. These oil solutions orsuspensions can also contain a long chain alcohol diluent or dispersant,carboxymethyl cellulose, or similar dispersing agents. Other commonlyused surfactants such as Tweens and Spans or other similar emulsifyingagents or bioavailability enhancers which are commonly used in themanufacture of pharmaceutically acceptable solid, liquid, or otherdosage forms can also be used for the purpose of formulation.

A composition for oral administration can be any orally acceptabledosage form including capsules, tablets, emulsions and aqueoussuspensions, dispersions, and solutions. In the case of tablets,commonly used carriers include lactose and corn starch. Lubricatingagents, such as magnesium stearate, are also typically added. For oraladministration in a capsule form, useful diluents include lactose anddried corn starch. When aqueous suspensions or emulsions areadministered orally, the active ingredient can be suspended or dissolvedin an oily phase combined with emulsifying or suspending agents. Ifdesired, certain sweetening, flavoring, or coloring agents can be added.

A nasal composition can be prepared according to techniques well knownin the art of pharmaceutical formulation. For example, such acomposition can be prepared as a solution in saline, employing benzylalcohol or other suitable preservatives, absorption promoters to enhancebioavailability, fluorocarbons, and/or other solubilizing or dispersingagents known in the art.

A pharmaceutical composition of this invention can also be administeredin the form of a suppository for rectal administration.

The carrier in a pharmaceutical composition must be “acceptable” in thesense that it is compatible with the active ingredient of thecomposition and, preferably, capable of stabilizing the activeingredient, and not deleterious to the subject to be treated. One ormore solubilizing agents can be utilized as pharmaceutical excipientsfor delivery of an active heterocyclic compound of this invention.Examples of other carriers include colloidal silicon oxide, magnesiumstearate, cellulose, sodium lauryl sulfate, and D&C Yellow #10.

The details of the invention are set forth in the description below.Other features, objects, and advantages of the invention will beapparent from the following detailed description of several embodiments,and also from the appending claims.

DETAILED DESCRIPTION

Disclosed first in detail are heterocyclic compounds of formula I:

A, Y¹, X¹, X², Y², X³, Y³, X⁴, and m are defined in the SUMMARY sectionabove.

In one embodiment, the compounds of formula I have Y¹ being phenylsubstituted with (R¹)_(n), 5-membered heteroaryl substituted with(R²)_(o), or 5-membered heterocycloalkenyl substituted with (R²)_(o), inwhich R¹ in (R¹)_(n) is, independently, F, Cl, Br, NO₂, CN, C₁-C₆ alkyl,C₁-C₆ haloalkyl, C₂-C₆ alkynyl, C₃-C₈ cycloalkyl, C₅-C₁₅heterocycloalkyl, aryl, heteroaryl, carbonyl, thionyl, iminyl, orspiroamino; and R² in (R²)_(o) is, independently, F, Cl, Br, NO₂, CN,amino, C₁-C₆ alkyl, C₁-C₆ haloalkyl, C₂-C₆ alkynyl, C₃-C₈ cycloalkyl,C₅-C₁₅ heterocycloalkyl, aryl, heteroaryl, carbonyl, thionyl, iminyl,spiroamino, or C₁-C₆ alkoxyl.

In one subset of this embodiment, the compounds of formula I have Y²being

Y³ being pyridyl, and R¹ being C₅-C₁₅ heterocycloalkyl.

In another embodiment, the compounds of formula I have Y² being

in which Z² is O or NRr.

In one subset of this embodiment, the compounds of formula I have Y²being

Y³ being pyridyl, and R¹ being amino.

In still another embodiment, the compounds of formula I are thosecovered by formula Ia:

in which R¹ is amino or C₅-C₁₅ heterocycloalkyl.

In one subset of this embodiment, the compounds of formula Ia have Y²being

Y³ being pyridyl; X³ being CH₂; X⁴ being CH₃, CH₂F, CHF₂, CF₃, or OCH₃,m being 1; and R¹ preferably being amino.

In another subset, the compounds have Y² being

Y³ being phenyl; X³ being CH₂; each of X⁴ being, independently, F, Cl,Br, CN, SO₂NH₂, CH₃, CH2F, CHF2, CF₃, OCF₃, C₁-C₆ alkoxyl, or amino; andm being 0-2.

In a third subset, the compounds have Y² being

Y³ being phenyl; X³ being deleted; each of X⁴ being, independently, F,Cl, Br, CN, SO₂NH₂, CH₃, CH₂F, CHF₂, CF₃, OCF₃, C₁-C₆ alkoxyl, or amino;and m being 0-2.

Formula Ia includes compounds in which Y² is

Y³ is phenyl or pyridyl, and X³ is CH₂. As an example, Y² is

Formula Ia further includes compounds in which Y² is

Y³ is phenyl or pyridyl, and X³ is CH₂. For instance, Y² is

In a fourth embodiment, the heterocyclic compounds of this invention arecovered by formula Ib:

A subset of compounds of formula Ib have Y¹ being

Y³ being phenyl or pyridyl; and X³ being deleted or CH₂.

In a different subset, compounds of formula Ib have Y¹ being

Y³ being phenyl or pyridyl; and X³ being deleted or CH₂, in which Q⁹ isN or CR⁵, R⁵ being H, F, Cl, Br, CN, amino, C₁-C₆ alkyl, C₁-C₆haloalkyl, or C₁-C₆ alkoxyl.

In another subset, compounds of this formula have Y¹ being

Y³ being phenyl or pyridyl; and X³ being deleted or CH₂.

Typically, the compounds of formula Ib have each of X⁴ being,independently, CH₃, CH₂F, CHF₂, CF₃, or OCH₃ and m being 0-2.

Exemplary compounds of formula I include, but are not limited to, thefollowing compounds:

The compounds of formula I can be prepared according to methods wellknown in the field. See, for example, R. Larock, Comprehensive OrganicTransformations (2nd Ed., VCH Publishers 1999); P. G. M. Wuts and T. W.Greene, Greene's Protective Groups in Organic Synthesis (4th Ed., JohnWiley and Sons 2007); L. Fieser and M. Fieser, Fieser and Fieser'sReagents for Organic Synthesis (John Wiley and Sons 1994); L. Paquette,ed., Encyclopedia of Reagents for Organic Synthesis (2nd ed., John Wileyand Sons 2009); and G. J. Yu et al., J. Med. Chem. 2008, 51, 6044-6054.

Also within the scope of this invention is a pharmaceutical compositioncontaining one or more of the heterocyclic compounds of formula I. Thepharmaceutical composition is used for treating a CSF1R modulatedcondition.

In certain embodiments, the pharmaceutical composition further containsone of the following therapeutic agents: an anti-proliferative agent, ananti-inflammatory agent, an immunomodulatory agent, and animmunosuppressive agent.

In other embodiments, the pharmaceutical composition further containsone of the following therapeutic agents: an alkylating agent, e.g.,adozelesin, altretamine, bizelesin, busulfan, carboplatin, carboquone,carmustine, chlorambucil, cisplatin, cyclophosphamide, dacarbazine,estramustine, fotemustine, hepsulfam, ifosfamide, improsulfan,irofulven, lomustine, mechlorethamine, melphalan, oxaliplatin,piposulfan, semustine, streptozocin, temozolomide, thiotepa, andtreosulfan; an antibody, e.g., alemtuzumab, bevacizumab, cetuximab,galiximab, gemtuzumab, nivolumab, panitumumab, pembrolizumab,pertuzumab, rituximab, tositumomab, trastuzumab, and 90 Y ibritumomabtiuxetan; a targeted signal transduction inhibitor, e.g., bortezomib,geldanamycin, and rapamycin; a kinase inhibitor, e.g., erlotinib,gefitinib, flavopiridol, imatinib mesylate, lapatinib, sorafenib,sunitinib malate, AEE-788, AG-013736, AMG 706, AMN107, BMS-354825,BMS-599626, 7-hydroxystaurosporine, vemurafenib, dabrafenib, trametinib,cobimetinib, selumetinib, and vatalanib; a taxane, e.g., DJ-927,docetaxel, TPI 287, paclitaxel, and DHA-paclitaxel; a retinoid, e.g.,alitretinoin, bexarotene, fenretinide, isotretinoin, and tretinoin; analkaloid, e.g., etoposide, homoharringtonine, teniposide, vinblastine,vincristine, vindesine, and vinorelbine; an antibiotic, e.g., bleomycin,dactinomycin, daunorubicin, doxorubicin, epirubicin, idarubicin,menogaril, mitomycin, mitoxantrone, neocarzinostatin, pentostatin, andplicamycin; an antiangiogenic agent, e.g., AE-941, ABT-510,2-methoxyestradiol, lenalidomide, and thalidomide; a topoisomeraseinhibitor, e.g., amsacrine, edotecarin, exatecan, irinotecan,7-ethyl-10-hydroxy-camptothecin, rubitecan, topotecan, and9-aminocamptothecin; an antimetabolite, e.g., azacitidine, capecitabine,cladribine, clofarabine, cytarabine, decitabine, floxuridine,fludarabine, 5-fluorouracil, ftorafur, gemcitabine, hydroxyurea,mercaptopurine, methotrexate, nelarabine, pemetrexed, raltitrexed,thioguanine, and trimetrexate; a hormone or hormone antagonist, e.g.,anastrozole, androgens, buserelin, diethylstilbestrol, exemestane,flutamide, fulvestrant, goserelin, idoxifene, letrozole, leuprolide,magestrol, raloxifene, tamoxifen, and toremifene; a biological responsemodifier, e.g., imiquimod, interferon-α, and interleukin-2; anindoleamine 2,3-dioxygenase inhibitor; a chemotherapeutic agent, e.g.,3-amino-2-carboxy aldehyde thiosemicarbazone, altrasentan,aminoglutethimide, anagrelide, asparaginase, bryostatin-1, cilengitide,elesclomol, eribulin mesylate, ixabepilone, lonidamine, masoprocol,mitoguanazone, oblimersen, sulindac, testolactone, and tiazofurin; amammalian target of rapamycin inhibitor; a phosphoinositide 3-kinaseinhibitor; a cyclin-dependent kinase 4 inhibitor; a protein kinase Binhibitor; a heat shock protein 90 inhibitor; a farnesyltransferaseinhibitor; an aromatase inhibitor (such as anastrozole, letrozole, andexemestane); a mitogen-activated protein kinase kinase inhibitor; atyrosine kinase inhibitor; an epidermal growth factor receptorinhibitor; a programmed cell death protein 1 inhibitor; a programmeddeath-ligand 1 inhibitor; or an interleukin 8 receptor beta inhibitor.

Still within the scope of this invention is a method of treating a CSF1Rmodulated condition using one or more of the above-describedheterocyclic compounds. For example, the condition can be a cancer,e.g., acute myeloid leukemia, bladder cancer, breast cancer, cervicalcancer, colon cancer, gastric cancer, gastrointestinal stromal tumor,glioblastoma multiforme, hepatocellular carcinoma, Hodgkin's lymphoma,kidney cancer, liver cancer, lung cancer, melanoma, metastatic tumor,ovarian cancer, pancreatic cancer, pigmented villondular synovitis,prostate cancer, tenosynovial giant cell tumors, endometrial cancer,multiple myeloma, myelocytic leukemia, bone cancer, renal cancer, braincancer, myeloproliferative disorder, esophageal cancer, squamous cellcarcinoma, uveal melanoma, follicular lymphoma, colorectal cancer, headand neck cancer, astrocytoma, and pulmonary adenocarcinoma; aninflammatory disorder or an autoimmune disease, e.g., psoriaticarthritis, arthritis, asthma, thyroiditis, glomerular nephritis,atherosclerosis, psoriasis, Sjogren's syndrome, rheumatoid arthritis,systemic lupus erythematosis, cutaneous lupus erythematosus, Crohn'sdisease, ulcerative colitis, type I diabetes, multiple sclerosis, humanimmunodeficiency virus encephalitis, Alzheimer's disease, amyotrophiclateral sclerosis, and epilepsy; or a bone disorder, e.g., osteoporosis,osteoarthritis, periodontitis, periprosthetic osteolysis, and Paget'sdisease.

Without further elaboration, it is believed that one skilled in the artcan, based on the above description, utilize the present invention toits fullest extent. The following specific examples are, therefore, tobe construed as merely illustrative, and not limitative of the remainderof the disclosure in any way whatsoever. All publications cited hereinare incorporated by reference in their entirety.

EXAMPLE 1: SYNTHESIS OF HETEROCYCLIC COMPOUNDS

Exemplary compounds of this invention, shown in Table 1 below, wereprepared by procedures shown in Scheme 1, Scheme 2, Scheme 3, or Scheme4. Table 1 includes mass spectral data for the compounds.

All chemicals and solvents were purchased from commercial suppliers andused as received. All reactions were carried out under an atmosphere ofdry nitrogen unless specified otherwise. Reactions were monitored bythin layer chromatography using Merck 60 F254 silica gel glass backedplates (5×10 cm) and zones were detected visually under ultravioletirradiation (254 nm) or by spraying with phosphomolybdic acid reagent(Aldrich) followed by heating at 80° C. Microwave reactions wereperformed in the CEM Discover SP System.

Flash column chromatography was performed by using Merck Kieselgel 60,No. 9385, 230-400 mesh ASTM silica gel as the stationary phase. Protonnuclear magnetic resonance (¹H NMR) spectra were measured on a VarianMercury-300 or Varian Mercury-400 spectrometer. Chemical shifts wererecorded in parts per million (ppm) on the delta (6) scale relative tothe resonance of the solvent peak. The following abbreviations were usedto describe coupling: s=singlet; d=doublet; t=triplet; q=quartet;quin=quintet; ABq=AB quartet; AA′XX′=second order AA′XX′ pattern;app.=apparent; br=broad; and m=multiplet.

Liquid chromatography mass spectrometry (LCMS) data was obtained with anAgilent MSD-1100 ESI-MS/MS, an Agilent 1200 series LC/MSD VL, or aWaters Acquity UPLC-ESI-MS/MS system.

Among the listed reagents and solvents in Scheme 1, SOCl₂ isthionylchloride, DMF is dimethylformamide, PhMe is toluene, tBuOK ispotassium tert-butoxide, THF is tetrahydrofuran, DMSO isdimethylsulfoxide, and CH₂Cl₂ is methylene chloride.

4-Chloro-7-fluoroquinazoline (B). To a suspension of7-fluoroquinazolin-4-ol compound A (6.32 g, 38.5 mmol) in dry PhMe (30mL), SOCl₂ (22 mL, 7.7 eq.) and DMF (2.6 mL) were added. The resultingmixture was refluxed for 10 h. The mixture was then cooled to roomtemperature, quenched with water (200 mL), and extracted with ethylacetate (EtOAc; 170 mL). The combined organic extracts were washed withwater (300 mL) and brine (30 mL), dried over sodium sulfate (NA₂SO₄),and concentrated to afford compound B (6.08 g, 86%) as a yellow solid.LCMS (ESI) m/z calculated for C₈H₄ClFN₂: 182, 184; found: 183, 185[M+H]⁺. ¹H-NMR (400 MHz, CDCl₃) 6 9.04 (s, 1H), 8.33 (dd, J=9.2 Hz,⁴J_(F,H)=6.0 Hz, 1H), 7.71 (dd, ³J_(F,H)=9.2 Hz, J=2.4 Hz, 1H), 7.52(ddd, J=9.2, 2.4 Hz, ³J_(F,H)=8.4 Hz, 1H).

4-((7-fluoroquinazolin-4-yl)oxy)aniline (C). A mixture of 4-aminophenolF (3.05 g, 28.0 mmol) and t-BuOK (3.14 g, 28.0 mmol) in dry THF (100 mL)was stirred for 20 min at 0° C. Subsequently, compound B (4.44 g, 24.3mmol) was added slowly in small portions. The reaction mixture was thenstirred for 3 h at 0° C., during which a suspension was formed.Afterwards, the suspension was filtered through a pad of celite, The padwas rinsed with THF. THF in the filtrate was removed by evaporation togive a crude residue. The residue was suspended in methanol (MeOH) andsonicated to form a solid. The solid was collected via filtration anddried to afford the titled product C (5.47 g, 88%) as an off-whitesolid. LCMS (ESI) m/z calculated for C₁₄H₁₀BrN₃O: 255; found: 256[M+H]⁺. ¹H-NMR (400 MHz, CDCl₃) δ 8.76 (s, 1H), 8.40 (dd, J=9.2 Hz,⁴J_(F,H)=6.0 Hz, 1H), 7.61 (dd, ³J_(F,H)=9.2 Hz, J=2.4 Hz, 1H), 7.40(ddd, J=9.2, 2.4 Hz, ³J_(F,H)=8.8 Hz, 1H), 7.04 (AA′XX′, J_(AX)=8.8 Hz,J_(AX′)=0 Hz, 2H), 6.77 (AA′XX′, J_(AX)=8.8 Hz, J_(AX′)=0 Hz, 2H), 3.71(br, 2H).

4-(4-aminophenoxy)-N,N-dimethylquinazolin-7-amine (D). To a solution ofcompound C (600 mg, 2.35 mmol) in DMSO (7.1 mL), 2 M dimethlyamine inTHF (3 eq., 3.53 mL) was added. The mixture was then irradiated inmicrowave for 10 min at 170° C. After compound C was consumed, thereaction mixture was diluted with EtOAc (30 mL) and 2% sodium carbonate(NA₂CO₃) solution (80 mL), and extracted with EtOAc (50 mL). Thecombined organic extracts were sequentially washed with 2% NA₂CO₃solution (160 mL), water (200 mL), and brine (20 mL), dried over NA₂SO₄,and concentrated. The crude residue was suspended in MeOH. Particles inthe supension were collected via filtration and dried to afford thetitled product D (207 mg, 32%) as a pale yellow solid. LCMS (ESI) m/zcalculated for C₁₆H₁₆N₄O: 280; found: 281 [M+H]⁺. ¹H-NMR (300 MHz,CDCl₃): δ 8.58 (s, 1H), 8.15 (d, J=9.2 Hz, 1H), 7.12 (dd, J=9.2 Hz,J=2.7 Hz, 1H), 7.03 (AA′XX′, J_(AX)=9.0 Hz, J_(AX′)=0 Hz, J_(AA′)=2.9Hz, J_(XX′)=2.9 Hz, 2H), 6.96 (d, J=2.7 Hz, 1H), 6.75 (AA′XX′,J_(AX)=9.0 Hz, J_(AX′)=0 Hz, J_(AA′)=2.9 Hz, J_(XX′)=2.9 Hz, 2H), 3.67(br, 2H), 3.15 (s, 6H).

1-(4-((7-(dimethylamino)quinazolin-4-yl)oxy)phenyl)-3-phenylurea(compound 4, E). To a solution of compound D (165 mg, 0.588 mmol) inCH₂Cl₂ (15 mL), compound H (108 μL, 0.883 mmol) was added. The mixturewas refluxed for 10 h, during which a suspension was formed. Particlesin the suspension were collected via filtration and rinsed with excessCH₂Cl₂ to afford the titled product E (214 mg, 91%) as a white solid.LCMS (ESI) m/z calculated for C₂₃H₂₁N₅O₂: 399; found: 400 [M+H]⁺. ¹H-NMR(400 MHz, DMSO-d₆): δ 8.77 (br s, 1H), 8.71 (br s, 1H), 8.45 (s, 1H),8.11 (d, J=9.6 Hz, 1H), 7.52 (AA′XX′, J_(AX)=8.8 Hz, J_(AX′)=0 Hz,J_(AA′)=2.6 Hz, J_(XX′)=2.6 Hz, 2H), 7.47 (d, J=5.6 Hz, 2H), 7.32-7.27(m, 3H), 7.19 (AA′XX′, J_(AX)=8.8 Hz, J_(AX′)=0 Hz, J_(AA′)=2.6 Hz,J_(XX′)=2.6 Hz, 2H), 6.97 (t, J=7.2 Hz, 1H), 6.88 (d, J=2.4 Hz, 1H),3.12 (s, 6H).

Compounds 1, 2, 3, 5, 6, 7, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19,20, 21, 52, and 106 were prepared in a manner similar to compound 4,with appropriate amines G and isocyanates H.

Among the listed reagents and solvents in Scheme 2, Py is pyridine, andEt₃N is triethylamine.

4-nitrophenyl ((6-(trifluoromethyl)pyridin-3-yl)methyl)carbamate (J). Toa −30° C. solution of (6-(trifluoromethyl)pyridin-3-yl)methanamine I(2.69 g, 15.3 mmol) and Py (1.23 mL, 15.3 mmol) in CH₂Cl₂ (50 mL),4-nitrophenyl chloroformate (3.85 g, 19.1 mmol) was slowly added. Thereaction mixture was stirred for 8 h and allowed to slowly warm to 0° C.Subsequently, water (50 mL) was added to the reaction mixture. Themixture was then stirred for 20 min, during which a suspension wasformed. Particles in the suspension were removed by filtering thereaction mixture through a pad of celite. The filtrate was washed with2% sodium bisulfate solution (20 mL), 2% sodium bicarbonate solution (40mL), water (20 mL), and brine (3 mL), dried over NA₂SO₄, andconcentrated. The crude residue was purified by column chromatography toafford the titled product J (3.33 g, quant.) as a white solid. LCMS(ESI) m/z calculated for C₁₄H₁₀F₃N₃O₄: 341; found: 342 [M+H]⁺. ¹H-NMR(400 MHz, CDCl₃): δ 8.73 (d, J=1.4 Hz, 1H), 8.26 (AA′XX′, J_(AX)=9.2 Hz,J_(AX′)=0 Hz, J_(AA′)=2.6 Hz, J_(xx′)=2.6 Hz, 2H), 7.91 (dd, J=8.0, 1.4Hz, 1H), 7.71 (d, J=8.0 Hz, 1H), 7.33 (AA′XX′, J_(AX)=9.2 Hz, J_(AX′)=0Hz, J_(AA′)=2.6 Hz, J_(XX′)=2.6 Hz, 2H), 5.68 (br t, 1H), 4.58 (d, J=6.4Hz, 2H).

1-(4-((7-(dimethylamino)quinazolin-4-yl)oxy)phenyl)-3-((6-(trifluoromethyl)pyridin-3-yl)methyl)urea(compound 27, K). To a solution of compound J (2.19 g, 6.43 mmol) andcompound D (1.06 g, 3.78 mmol) in CH₂Cl₂ (95 mL), Et₃N (1.05 mL) wasadded. The reaction mixture was refluxed for 4 d, during which asuspension was formed. Particles in the suspension were collected viafiltration and rinsed with excess CH₂Cl₂ to afford the titled product K(1.33 g, 73%) as a white solid. LCMS (ESI) m/z calculated forC₂₄H₂₁F₃N₆O₂: 482; found: 483 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆): δ 8.83(s, 1H), 8.72 (d, J=1.2 Hz, 1H), 8.43 (s, 1H), 8.10 (d, J=9.2 Hz, 1H),8.00 (dd, J=8.4, 1.2 Hz, 1H), 7.89 (d, J=8.4 Hz, 1H), 7.47 (AA′XX′,J_(AX)=8.8 Hz, J_(AX′)=0 Hz, J_(AA′)=2.6 Hz, J_(XX′)=2.6 Hz, 2H), 7.29(d, J=9.2, 2.4 Hz, 1H), 7.13 (AA′XX′, J_(AX)=8.8 Hz, J_(AX′)=0 Hz,J_(AA′)=2.6 Hz, J_(XX′)=2.6 Hz, 2H), 6.87 (d, J=2.4 Hz, 1H), 6.84 (t,J=6.0 Hz, 1H), 4.44 (d, J=6.0 Hz, 2H), 3.12 (s, 6H).

1-(4-((7-(dimethylamino)quinazolin-4-yl)oxy)phenyl)-3-((6-methylpyridin-3-yl)methyl)urea(compound 65, K) LCMS (ESI) m/z calculated for C₂₄H₂₁F₃N₆O₂: 428; found:429 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 8.68 (s, 1H), 8.43 (s, 1H), 8.39(d, J=2.4 Hz, 1H), 8.10 (d, J=9.2 Hz, 1H), 7.61 (dd, J=8.0, 2.4 Hz, 1H),7.46 (AA′XX′, J_(AX)=8.8 Hz, 2H), 7.29 (dd, J=9.2, 2.4 Hz, 1H), 7.22 (d,J=8.0 Hz, 1H), 7.13 (AA′XX′, J_(AX)=8.8 Hz, 2H), 6.87 (d, J=2.4 Hz, 1H),6.67 (t, J=6.0 Hz, 1H), 4.28 (d, J=6.0 Hz, 1H), 3.12 (s, 6H), 2.44 (s,3H).

1-(4-((7-(dimethylamino)quinazolin-4-yl)oxy)phenyl)-3-((6-methylpyridin-3-yl)methyl)urea(compound 66, K) LCMS (ESI) m/z calculated for C₂₄H₂₁F₃N₆O₂: 444; found:445 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 8.68 (s, 1H), 8.43 (s, 1H), 8.39(d, J=2.4 Hz, 1H), 8.10 (d, J=9.2 Hz, 1H), 7.61 (dd, J=8.0, 2.4 Hz, 1H),7.46 (AA′XX′, J_(AX)=8.8 Hz, 2H), 7.29 (dd, J=9.2, 2.4 Hz, 1H), 7.22 (d,J=8.0 Hz, 1H), 7.13 (AA′XX′, J_(AX)=8.8 Hz, 2H), 6.87 (d, J=2.4 Hz, 1H),6.67 (t, J=6.0 Hz, 1H), 4.28 (d, J=6.0 Hz, 1H), 3.12 (s, 6H), 2.44 (s,3H).

Compounds 23, 24, 25, 28, 30, 33, 36, 39, 40, 42, 43, 44, 49, 50, 51,54, 55, 56, 57, 58, 59, 60, 63, 65, 66, 87, 90, 91, 92, 96, and 101 wereprepared in a manner similar to compound 27 with appropriate amines Hand anilines D

Among the listed reagents, solvents and catalysts in Scheme 3, SOCl₂ isthionylchloride, RuPhos Pd G3 is(2-dicyclohexylphosphino-2′,6′-diisopropoxy-1,1′-biphenyl)[2-2′-amino-1,1′-biphenyl]]palladium(II) methanesulfonate, RuPhos is2-dicyclohexylphosphino-2′,6′-diisopropoxybiphenyl, and Cs₂CO₃ is cesiumcarbonate.

4-Chloro-7-bromoquinazoline (M). To a suspension of7-bromoquinazolin-4-ol L (1.17 g, 5.20 mmol) in dry PhMe (6 mL), SOCl₂(6 mL) and DMF (0.6 mL) were added. The resulting mixture was stirred at95° C. for 9 h, cooled to room temperature, quenched with water (100mL), and extracted with EtOAc (70 mL). The combined organic extractswere washed with water (200 mL) and brine (5 mL), dried over NA₂SO₄, andconcentrated to afford the titled product M (1.26 g, 99%) as a yellowsolid. LCMS (ESI) m/z calculated for C₈H₄BrClN₂: 242, 244, 246; found:243, 245, 247 [M+H]⁺. ¹H-NMR (300 MHz, CDCl₃): δ 9.05 (s, 1H), 8.28 (d,J=1.8 Hz, 1H), 8.15 (d, J=9.0 Hz, 1H), 7.84 (dd, J=9.0, 1.8 Hz, 1H).

4-((7-Bromoquinazolin-4-yl)oxy)aniline (N). A mixture of 4-aminophenol(0.678 g, 6.21 mmol) and t-BuOK (0.668 g, 5.95 mmol) in dry THF (15 mL)at 0° C. was stirred for 20 min. Subsequently,4-chloro-7-bromoquinazoline M (1.26 g, 5.17 mmol) was added slowly insmall portions. The reaction mixture was then stirred for 3 h at 0° C.,during which a suspension was formed. The suspension was filteredthrough a pad of celite. The pad was rinsed with THF (20 mL), and theresulting filtrate was evaporated to give a crude solid. The crude solidwas suspended in MeOH (6 mL) and sonicated. Particles in the suspensionwere collected via filtration and dried to afford the titled product N(1.461 g, 89%) as an off-white solid. LCMS (ESI) m/z calculated forC₁₄H₁₀BrN₃O: 315, 317; found: 316, 318 [M+H]⁺.

1-(4-((7-Bromoquinazolin-4-yl)oxy)phenyl)-3-((6-(trifluoromethyl)pyridin-3-yl)methyl)urea(O; compound 31). To a solution of4-((7-Bromoquinazolin-4-yl)oxy)aniline N (305 mg, 0.964 mmol) andcompound Q (461 mg, 1.35 mmol) in CH₂Cl₂ (6 mL), Et₃N (0.27 mL) wasadded. The resulting mixture was refluxed for 1.5 d, during which asuspension was formed. Particles in the suspension was collected viafiltration and rinsed with excess CH₂Cl₂ to afford the titled product O(462 mg, 93%) as a white solid. LCMS (ESI) m/z calculated forC₂₂H₁₅BrF₃N₅O₂: 517, 519; found: 518, 520 [M+H]⁺. ¹H NMR (400 MHz,DMSO-d6): δ 8.87 (s, 1H), 8.74 (s, 1H), 8.72 (br s, 1H), 8.30 (d, J=8.8Hz, 1H), 8.24 (d, J=2.0 Hz, 1H), 8.00 (dd, J=8.0, 1.6 Hz, 1H), 7.93 (dd,J=8.8, 2.0 Hz, 1H), 7.90 (d, J=8.0 Hz, 1H), 7.50 (AA′XX′, J_(AX)=9.0 Hz,J_(AX′)=0 Hz, 2H), 7.21 (AA′XX′, J_(AX)=9.0 Hz, J_(AX′)=0 Hz, 2H), 6.86(t, J=6.0 Hz, 1H), 4.44 (d, J=6.0 Hz, 2H).

1-(4-((7-(3,5-Dimethylpiperazin-1-yl)quinazolin-4-yl)oxy)phenyl)-3-((6-(trifluoromethyl)pyridin-3-yl)methyl)urea(P; compound 34). Freshly-distilled THF (4 mL) was purged with argon for20 min, after which compound O (50 mg, 0.097 mmol), Cs₂CO₃ (47 mg, 0.15mmol), cis-2,6-dimethylpiperazine (17 mg, 0.15 mmol), Ruphos (4.1 mg,0.009 mmol), and Ruphos Pd G3 (4.1 mg, 0.005 mmol) were added to form asuspension. The resulting mixture was stirred under argon for 5 min.Subsequently, the reaction mixture was refluxed overnight. The resultingpale-yellow solution was filtered through a pad of celite and thefiltrate was concentrated to give a residue. The residue was purified bycolumn chromatography to afford the titled product P (49 mg, 92%) as awhite solid. LCMS (ESI) m/z calculated for C₂₈H₂₈F₃N₇O₂: 551; found: 552[M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆): δ 8.85 (s, 1H), 8.72 (d, J=1.6 Hz,1H), 8.46 (s, 1H), 8.09 (d, J=9.2 Hz, 1H), 8.00 (dd, J=8.0, 1.6 Hz, 1H),7.90 (d, J=8.0 Hz, 1H), 7.51 (dd, J=9.2, 2.4 Hz, 1H), 7.47 (AA′XX′,J_(AX)=9.2 Hz, J_(AX′)=0 Hz, 2H), 7.17-7.13 (m, 3H), 6.85 (t, J=5.6 Hz,1H), 4.44 (d, J=5.6 Hz, 2H), 3.91-3.88 (m, 2H), 2.87-2.82 (m, 2H),2.39-2.33 (m, 2H), 1.06 (d, J=6.4 Hz, 6H).

1-(4-((7-(4-hydroxypiperidin-1-yl)quinazolin-4-yl)oxy)phenyl)-3-((6-(trifluoromethyl)pyridin-3-yl)methyl)urea(P; compound 30). Similar to compound 34, compound 30 was prepared fromcompound 31 and was obtained as a white solid. LCMS (ESI) m/z calculatedfor C₂₈H₂₈F₃N₇O₂: 538; found: 539 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆): δ8.82 (br s, 1H), 8.72 (d, J=1.2 Hz, 1H), 8.45 (s, 1H), 8.08 (d, J=9.2Hz, 1H), 8.00 (dd, J=8.0, 1.2 Hz, 1H), 7.89 (d, J=8.0 Hz, 1H), 7.51-7.45(m, 3H), 7.15-7.11 (m, 3H), 6.84 (t, J=6.0 Hz, 1H), 4.76 (d, J=4.0 Hz,2H), 4.44 (d, J=6.0 Hz, 2H), 3.87-3.81 (m, 2H), 3.77-3.72 (m, 1H),3.21-3.14 (m, 1H), 1.87-1.83 (m, 2H), 1.51-1.43 (m, 2H).

Compounds 24, 96, 97, 98, 99, 100 were prepared analogously to compound34.

Among the listed reagents, solvents, and catalysts listed in Scheme 3,POCl₃ is phosphoryl chloride, CuI is copper iodide, 1,10-phen is1,10-phenanthroline, K₂CO₃ is potassium carbonate, and tBuOH istert-butanol.

7-(Dimethylamino)quinazolin-4(3H)-one (R). A solution of7-fluoroquinazolin-4(3H)-one A (1.65 g, 10.1 mmol) and dimethylamine (2Min THF, 20 mL, 40.2 mmol) in 2-methoxyethanol (60 mL) in a 250 mL sealedtube was stirred at 130° C. for 1 d, during which a solid was formed.Subsequently, the reaction mixture was cooled to room temperature andconcentrated under reduced pressure. The resulting residue was suspendedin MeOH (8 mL) and filtered to afford the titled product R (1.5 g, 79%)as a brown solid. LCMS (ESI) m/z calculated for C₁₀H₁₁N₃O: 189; found:190 [M+H]⁺. ¹H NMR (300 MHz, DMSO-d₆): δ 11.69 (br, 1H), 7.91 (s, 1H),7.88 (d, J=9.0 Hz, 1H), 6.93 (dd, J=9.0, 2.4 Hz, 1H), 6.98 (d, J=2.4 Hz,1H), 3.04 (s, 6H).

4-Chloro-N,N-dimethylquinazolin-7-amine (S). To a suspension of7-(dimethylamino)quinazolin-4(3H)-one R (910 mg, 4.81 mmol) in dry PhMe(10 mL), POCl₃ (4 mL) was added, after which the resulting mixture wasstirred at 95° C. for 12 h. The reaction mixture was the cooled to 0°C., quenched with ice water (100 mL), and neutralized with saturatedsodium carbonate solution until the color of the solution turned frombright orange to pale yellow. Subsequently, the reaction mixture wasextracted with EtOAc (170 mL), and the organic extract was washed withwater (200 mL) and brine (20 mL), dried over NA₂SO₄, and concentrated toafford the titled product S (948 mg, 95%) as a yellow solid. LCMS (ESI)m/z calculated for C₁₀H₁₀ClN₃: 207, 209; found: 208, 210 [M+H]⁺. ¹H-NMR(300 MHz, CDCl₃): δ 8.77 (s, 1H), 8.03 (d, J=9.3 Hz, 1H), 7.20 (dd,J=9.3, 2.7 Hz, 1H), 6.96 (d, J=2.7 Hz, 1H), 3.18 (s, 1H).

4-((1H-indol-5-yl)oxy)-N,N-dimethylquinazolin-7-amine (T). DMF (4 mL)was purged with argon. Subsequently,4-Chloro-N,N-dimethylquinazolin-7-amine S (322 mg, 1.55 mmol),5-hydroxyindole (413 mg, 3.10 mmol), K₂CO₃ (429 mg, 3.10 mmol), CuI(29.5 mg, 0.155 mmol), and 1,10-phen (27.9 mg, 0.155 mmol) were added toform a reaction mixture, which was stirred at 90° C. for 3 h underargon. The reaction mixture was then diluted with EtOAc (15 mL) andwater (15 mL), and filtered through a pad of celite. The filtrate wasextracted with EtOAc (60 mL), and the organic extract was washed withwater (120 mL) and brine (5 mL), dried over NA₂SO₄, and concentrated.The residue was suspended in MeOH and particles were collected byfiltration to afford the titled product T (381 mg, 81%) as a whitesolid. LCMS (ESI) m/z calculated for C₁₈H₁₆N₄O: 304; found: 305 [M+H]⁺.

5-((7-(dimethylamino)quinazolin-4-yl)oxy)-N-((6-(trifluoromethyl)pyridin-3-yl)methyl)-1H-indole-1-carboxamide(U; compound 67). To a solution of4-(4-amino-3-methoxyphenoxy)-N,N-dimethylquinazolin-7-amine T (170 mg,0.95 mmol) in dry THF (5 mL), NaH (60% in oil, 112 mg, 2.79 mmol) wasadded. The resulting mixture was stirred at room temperature for 20 min.Subsequently, the mixture was cooled to −60° C., and 4-nitrophenyl((6-(trifluoromethyl)pyridin-3-yl)methyl)carbamate V (574 mg, 1.68 mmol)was added in 3 portions. The mixture was then slowly warmed to 0° C.,stirred for 1 h, and quenched with saturated ammonium chloride solution.The solvent was removed under reduced pressure and the resulting residuewas diluted with EtOAc (35 mL), washed with water (80 mL) and brine (3mL), dried over NA₂SO₄, and concentrated. The crude residue was purifiedby column chromatography to afford the titled product U (171 mg, 65%) asa white solid. LCMS (ESI) m/z calculated for C₂₆H₂₁F₃N₆O₂: 506; found:507 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆): δ 8.94 (t, J=5.6 Hz, 1H), 8.82(d, J=1.6 Hz, 1H), 8.41 (s, 1H), 8.27 (d, J=8.8 Hz, 1H), 8.15 (d, J=9.2Hz, 1H), 8.10 (dd, J=8.0, 1.6 Hz, 1H), 7.96 (d, J=3.6 Hz, 1H), 7.92 (d,J=8.0 Hz, 1H), 7.50 (d, J=2.4 Hz, 1H), 7.31 (dd, J=9.2, 2.4 Hz, 1H) 7.16(dd, J=8.8, 2.4 Hz, 1H), 6.88 (d, J=2.4 Hz, 1H), 6.74 (d, J=3.6 Hz, 1H),4.64 (d, J=5.6 Hz, 2H), 3.13 (s, 6H).

Compounds 29, 35, 38, 64, 68, 69, 83, and 89 were prepared in a mannersimilar to compound 67.

TABLE 1 Exemplary heterocyclic compounds Com- Calculated Mass poundStructure Mass [M + H]⁺  1

455 456  2

469 470  3

454 455  4

399 400  5

469 470  6

469 470  7

434 435  8

374 375  9

425 426  10

469 470  11

482 483  12

443 444  13

467 468  14

417 418  15

433 434  16

433 434  17

503 504  18

417 418  19

433 434  20

503 504  21

429 430  22

415 416  23

565 566  24

552 553  25

516 517  26

498 499  27

482 483  28

537 538  29

510 511  30

538 539  31

517 518  32

457 458  33

516 517  34

551 552  35

510 511  36

500 501  37

498 499  38

496 497  39

526 527  40

512 513  41

501 502  42

512 513  43

552 553  44

539 540  45

595 596  46

551 552  47

496 497  48

495 496  49

492 493  50

419 420  51

413 414  52

449 450  53

467 468  54

414 415  55

473 474  56

481 482  57

497 498  58

427 428  59

443 444  60

438 439  61

499 500  62

503 504  63

403 404  64

457 458  65

428 429  66

444 445  67

506 507  68

507 508  69

507 508  71

398 399  72

420 421  73

481 482  74

495 496  75

506 507  76

471 472  77

558 559  78

503 504  79

481 482  80

506 507  81

464 465  82

483 484  83

507 508  84

417 418  85

519 520  86

519 520  87

409 410  88

507 508  89

507 508  90

393 394  91

434 435  92

421 422  93

428 429  94

429 430  95

443 444  96

551 552  97

565 566  98

579 580  99

524 525 100

551 552 101

420 421 102

480 481 103

496 497 104

463 465 464 466 105

463 465 464 466 106

417 418 107

385 386 108

581 582

EXAMPLE 2: CSF1R KINASE ASSAY

A study was conducted to test certain compounds described in EXAMPLE 1in inhibiting CSF1R kinase activity. Results from this study are shownin Table 2 below (see column 2).

Activity of CSF1R kinase was determined using a CSF1R Kinase-Glo assay.Recombinant N-terminal GST-CSF1R (CSF1R residues L534-C972) containingthe CSFIR kinase domain was expressed in Sf9 insect cells and purified.The kinase assay was carried out in 96-well plates at 30° C. for 180 minwith the tested compounds in a final volume of 50 μl including thefollowing components: 25 mM Tris-HCl pH 7.4, 4 mM MnCl₂, 10 mM MgCl₂,0.01% BSA, 0.5 mM NA₃VO₄, 0.02% Triton X-100, 40 μM ATP, 2 mM DTT and 20μM poly(Glu,Tyr) 4:1 peptide, and 600 ng recombinant GST-CSF1R.Following incubation, 50 μl Kinase-Glo Plus Reagent (Promega, Madison,Wis., USA) was added and the mixture was incubated at 25° C. for 20 min.A 70-μL aliquot of each reaction mixture was transferred to a blackmicrotiter plate and luminescence was measured on Wallac Vector 1420multilabel counter (PerkinElmer, Shelton, Conn., USA). Percentageinhibition values were obtained by comparison of reaction rate with therates in control wells (i.e., in the absence of test compound). IC₅₀values were calculated from a series of percentage inhibition valuesdetermined at a range of test compound concentrations using GraphPadPrism version 6 software (GraphPad, San Diego, Calif., USA).

EXAMPLE 3: CELLULAR PROLIFERATION ASSAYS

Studies were conducted to evaluate in vivo anticancer activity ofcertain compounds described in EXAMPLE 1 using3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium)(MTS) cell viability assays. Results from these studies are shown inTable 2 below (see columns 3 and 4).

Cell line and culture:

The cell lines M-NFS-60 (ATCC®CRL-1838™) and BaF3-CSF1R-1600 wereobtained from American Type Culture Collection (ATCC, Manassas, Va.,USA). The stable BaF3-CSF1R-1600 cell line expresses ETV6-CSF1R fusionprotein consisting of N-terminal ETS-variant gene 6 protein (ETV6residues M1-G337) and CSF1R tyrosine kinase (CSF1R residues L533-C972).The M-NSF-60 and BaF3-CSF1R-1600 cells were cultured in RPMI1640 mediumsupplemented with 10% fetal bovine serum, 0.05 mM 2-ME, 10 U/mlpenicillin, and 10 g/ml streptomycin at 37° C. and 5% CO₂.

MTS cell viability assays:

M-NFS-60 and BaF3-CSF1R-1600 cells were seeded in 96-well plates at adensity of 10000 cells/100 μl and 8000 cells/100 μl per well,respectively, for 16 h and treated with vehicle or variousconcentrations of test compounds in medium for 72 h. Viable cells werequantified using the MTS method (Promega, Madison, Wis., USA) accordingto manufacturer's recommended protocol. The results were determined bymeasuring absorbance at 490 nm using a plate reader (Victor 2). The GI50value was defined as the amount of compound that caused 50% reduction incell viability in comparison with DMSO-treated (vehicle) control and wascalculated using Prism GraphPad Prism version 6 software (GraphPad).

TABLE 2 Inhibitory activity of heterocyclic compounds in a CSF1R kinaseassay and in M-NFS-60 and BaF3-CSF1R cell assays Enzymatic AssayCellular CSF1R Proliferation Assay enzyme M-NFS-60 (nM) BaF3-CSF1RCompound IC₅₀, (nM) GI₅₀ (nM) GI₅₀ (nM)  1 +++ ++ ++  2 +++ +++ +++  3+++ +++ +++  4 +++ +++ +++  5 +++ ++  6 +++  7 +  8 ++  9 +++ +++ +++ 10 +++ ++ ++  11 +++ +++ +++  12 +++  13 ++  14 ++++ ++ ++++  15 ++++++ +++  16 ++++ +++ +++  17 ++++ +++ +++  18 ++++ ++ +++  19 ++++ +++++++  20 ++++ +++ +++  21 ++++ +++ +++  22 ++  23 +++ +++ +++  24 +++++++ +++  25 +++ + ++  26 +++ ++ +++  27 ++++ +++ +++  28 +++ +++ ++++ 29 ++  30 ++++ +++ +++  31 ++ + ++  32 +++ + ++  33 +++ ++ ++  34 ++++++ ++++  35 +  36 +++ ++  37 ++  38 +  39 ++++ ++  40 +++ ++ +++  41 +++++  42 ++++ ++ +++  43 +++ ++ ++  44 ++++ ++ +++  45 ++++ ++ +++  47 + 49 ++++ + ++  50 +++  51 ++++ +++ +++  52 ++  53 ++  54 ++  55 +++  56+++ ++ +++  57 ++++ ++ ++  58 ++++ ++ +++  59 ++++ +++ +++  60 +++  61++  62 ++++ ++  63 +++ ++ ++  64 +  65 ++++ +++ +++  66 ++++ +++ +++  67+++ ++ +++  68 +++ ++ +++  69 +  71 ++  72 +  73 +++  75 +  77 ++  79++++  83 ++  84 ++  85 +++  87 +  89 ++  90 ++  93 +++  97 +++ +++ +++ 98 ++++ +++ ++++  99 ++++ +++ +++ 100 ++++ +++ +++ 101 +++ ++ ++ 102+++ + ++ 104 +++ ++ +++ 105 +++ +++ +++ 106 ++++ ++ + 107 +++ 108 ++++++ +++ ^(a) ++++: IC₅₀ < 20 nM; +++: 20 nM < IC₅₀ < 100 nM; ++: 100 nM< IC₅₀ < 1000 nM; +: ^(b) ++++: GI₅₀ < 20 nM; +++: 20 nM < GI₅₀ < 100nM; ++: 100 nM < GI₅₀ < 1000 nM; +:

EXAMPLE 4. KINASE SELECTIVITY PROFILE

A study was conducted to determine kinase selectivity of compounds 27and 67. More specifically, each compound was tested for inhibitoryactivity of CSF1R kinase, as compared to that of seven other kinases,i.e., Aurora A, Aurora B, tyrosine-protein kinase Kit (c-Kit), fms-liketyrosine kinase 3 (FLT3), platelet-derived growth factor receptor(PDGFR) A, PDGFR B, and discoidin domain receptor tyrosine kinase 1(DDR1). Results from this study are shown in Table 3 below.

In vitro profiling of the kinase panel was performed at Reaction BiologyCorporation (www.reactionbiology.com, Malvern, Pa.) using the “HotSpot”assay platform. Briefly, specific kinase/substrate pairs along withrequired cofactors were prepared in a reaction buffer containing 20 mMHepes pH 7.5, 10 mM MgCl₂, 1 mM EGTA, 0.02% Brij35, 0.02 mg/ml BSA, 0.1mM NA₃VO₄, 2 mM DTT, and 1% DMSO. Compound 27 or 67 was added. 20minutes later, a mixture of ATP (Sigma, St. Louis Mo.) and ³³P ATP(Perkin Elmer, Waltham Mass.) was added to the resulting reactionsolution to give a final concentration of 10 μM. The reaction wascarried out at room temperature for 120 min., followed by spotting thereaction solution onto P81 ion exchange filter paper (Whatman Inc.,Piscataway, N.J.). Unbound phosphate was removed by extensive washingfilters in 0.75% phosphoric acid. After subtracting the backgroundderived from a control reaction containing an inactive enzyme, kinaseactivity was determined as the percentage of the remaining kinaseactivity in a test sample compared to a vehicle (DMSO) reaction. An IC₅₀value and a dose-response curve for each compound against each kinasewere obtained using Prism (Graph Pad Software). Selectivity, expressedas an IC₅₀ ratio, was determined by dividing the IC₅₀ value of a kinase,e.g., Aurora A, by that of CSF1R.

TABLE 3 Kinase selectivity profile Kinase IC₅₀ (nM) Compound CSF1RAurora A Aurora B c-Kit FLT3 PDGFR A PDGFR B DDR1 27 0.534 ++++ +++ +++++ ++ ++ +++ 67 5.69 +++ +++ +++ +++ ++ +++ + ++++: IC₅₀ ratio > 1000;+++: 1000 > IC₅₀ ratio > 100; ++: 100 > IC₅₀ ratio > 10; +: 10 > IC₅₀ratio > 1

OTHER EMBODIMENTS

All of the features disclosed in this specification may be combined inany combination. Each feature disclosed in this specification may bereplaced by an alternative feature serving the same, equivalent, orsimilar purpose. Thus, unless expressly stated otherwise, each featuredisclosed is only an example of a generic series of equivalent orsimilar features.

Further, from the above description, one skilled in the art can easilyascertain the essential characteristics of the present invention, andwithout departing from the spirit and scope thereof, can make variouschanges and modifications of the invention to adapt it to various usagesand conditions. Thus, other embodiments are also within the claims.

1. A compound of formula I:

wherein A is H, C₁-C₆ alkyl, or ORr, Rr being H or C₁₋₆ alkyl; Y¹ isphenyl substituted with (R¹)_(n), 5-membered heteroaryl substituted with(R²)_(o), 5-membered heterocycloalkenyl substituted with (R²)_(o), oralkenyl substituted with (R²)_(o), in which R¹ in (R¹)_(n), n being 0-4,is, independently, F, Cl, Br, NO₂, CN, amino, C₁-C₆ alkyl, C₁-C₆haloalkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₈ cycloalkyl, C₅-C₁₅heterocycloalkyl, aryl, heteroaryl, carbonyl, thionyl, iminyl, orspiroamino; and R² in (R²)_(o), o being 0-5, is, independently, F, Cl,Br, NO₂, CN, amino, C₁-C₆ alkyl, C₁-C₆ haloalkyl, C₂-C₆ alkenyl, C₂-C₆alkynyl, C₃-C₈ cycloalkyl, C₅-C₁₅ heterocycloalkyl, aryl, heteroaryl,carbonyl, thionyl, iminyl, spiroamino, or C₁-C₆ alkoxyl; X¹ is N or CR³,R³ being H, F, Cl, Br, CN, C₁-C₆ alkyl, C₁-C₆ haloalkyl, or C₁-C₆alkoxyl; X² is O, S, NH, or CH₂; Y²is

in which each of Q¹, Q², Q³, Q⁴, Q⁵, Q⁶, Q⁷, and Q⁸ is, independently, Nor CR⁴, R⁴ being H, F, Cl, Br, CN, amino, C₁-C₆ alkyl, C₁-C₆ haloalkyl,or C₁-C₆ alkoxyl; Z¹ is O, S, or NRr; Z² is O, S, or NRr; and G and Hare, respectively, C or N and N or C; X³ is deleted, CH₂, (CH₂)₂, orCH(C≡CH); Y³ is C₁-C₆ alkyl, aryl, heteroaryl, C₃-C₈ cycloalkyl, orC₅-C₆ heterocycloalkyl having one heteroatom, in which the oneheteroatom is O or N; and X⁴ in (X⁴)_(m), m being 0-5, is,independently, F, Cl, Br, CN, SO₂NH₂, amino, C₁-C₆ alkyl, C₁-C₆haloalkyl, or C₁-C₆ alkoxyl.
 2. The compound of claim 1, wherein Y¹ isphenyl substituted with (R¹)_(n), 5-membered heteroaryl substituted with(R²)_(o), or 5-membered heterocycloalkenyl substituted with (R²)_(o), inwhich R¹ in (R¹)_(n) is, independently, F, Cl, Br, NO₂, CN, C₁-C₆ alkyl,C₁-C₆ haloalkyl, C₂-C₆ alkynyl, C₃-C₈ cycloalkyl, C₅-C₁₅heterocycloalkyl, aryl, heteroaryl, carbonyl, thionyl, iminyl, orspiroamino; and R² in (R²)_(o) is, independently, F, Cl, Br, NO₂, CN,amino, C₁-C₆ alkyl, C₁-C₆ haloalkyl, C₂-C₆ alkynyl, C₃-C₈ cycloalkyl,C₅-C₁₅ heterocycloalkyl, aryl, heteroaryl, carbonyl, thionyl, iminyl,spiroamino, or C₁-C₆ alkoxyl.
 3. The compound of claim 2, wherein Y² is

Y³ is pyridyl, and R¹ is C₅-C₁₅ heterocycloalkyl.
 4. The compound ofclaim 1, wherein Y² is

in which Z² is O or NRr.
 5. The compound of claim 4, wherein Y² is

Y³ is pyridyl, and R¹ is amino.
 6. The compound of claim 1, wherein thecompound is of formula Ia:

in which R¹ is amino or C₅-C₁₅ heterocycloalkyl.
 7. The compound ofclaim 6, wherein Y² is

Y³ is pyridyl; X³ is CH₂; X⁴ is CH₃, CH₂F, CHF₂, CF₃, or OCH₃; and mis
 1. 8. The compound of claim 6, wherein Y² is

Y³ is phenyl; X³ is CH₂; each of X⁴ is, independently, F, Cl, Br, CN,SO₂NH₂, CH₃, CH₂F, CHF₂, CF₃, OCF₃, C₁-C₆ alkoxyl, or amino; and m is0-2.
 9. The compound of claim 6, wherein Y² is

Y³ is phenyl; X³ is deleted; each of X⁴ is, independently, F, Cl, Br,CN, SO₂NH₂, CH₃, CH₂F, CHF₂, CF₃, OCF₃, C₁-C₆ alkoxyl, or amino; and mis 0-2.
 10. The compound of claim 7, wherein R¹ is amino.
 11. Thecompound of claim 6, wherein Y² is

Y³ is phenyl or pyridyl; and X³ is CH₂.
 12. The compound of claim 11,wherein Y² is


13. The compound of claim 6, wherein Y² is

Y³ is phenyl or pyridyl; and X³ is CH₂.
 14. The compound of claim 13,wherein Y² is


15. The compound of claim 1, wherein the compound is of formula Ib:


16. The compound of claim 15, wherein Y¹ is

Y³ is phenyl or pyridyl; and X³ is deleted or CH₂.
 17. The compound ofclaim 16, wherein each of X⁴ is, independently, CH₃, CH₂F, CHF₂, CF₃, orOCH3 and m is 0-2.
 18. The compound of claim 15, wherein Y¹ is

Y³ is phenyl or pyridyl; and X³ is deleted or CH₂, in which Q⁹ is N orCR⁵, R⁵ being H, F, Cl, Br, CN, amino, C₁-C₆ alkyl, C₁-C₆ haloalkyl, orC₁-C₆ alkoxyl.
 19. The compound of claim 18, wherein each of X⁴ is,independently, CH₃, CH₂F, CHF₂, CF₃, or OCH3 and m is 0-2.
 20. Thecompound of claim 15, wherein Y¹ is

Y³ is phenyl or pyridyl; and X³ is deleted or CH₂.
 21. The compound ofclaim 20, wherein each of X⁴ is, independently, CH₃, CH₂F, CHF₂, CF₃, orOCH₃ and m is 0-2.
 22. The compound of claim 1, wherein the compound isone of the following compounds:


23. The compound of claim 1, wherein the compound is


24. The compound of claim 1, wherein the compound is

25-31. (canceled)